Li Yong, Zhu Xinghua, Zhang Can, Yin Yi, Chen Lei, Liu Yushan, He Aiqin, Xia Fei
Department of Gynecological Oncology, Nantong Tumor Hospital, Affiliated Tumor Hospital of Nantong University, Nantong, Jiangsu, China.
Department of Gynecology and Obstetrics, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
Histol Histopathol. 2023 Dec;38(12):1487-1498. doi: 10.14670/HH-18-620. Epub 2023 Jan 9.
Overexpressed long noncoding RNA FTX is associated with low survival rate of epithelial ovarian cancer (EOC) patients, and enhances tumor infiltration. Thus, we aim to illuminate the undefined underlying mechanisms. Real-time quantitative polymerase chain reaction was applied to detect the expressions of FTX, miR-7515, miR-342-3p, miR-940, miR-150-5p, miR-205-5p and tumor protein D52 (TPD52). Cell counting kit-8 and transwell assays were utilized to explore the cell viability, migration or invasion of EOC cells. Western blot was conducted to measure the expressions of E-cadherin, N-cadherin, Met, phosphorylated (p)-Met, Akt, p-Akt, mTOR and p-mTOR. LncBase and TargetScan predicted the binding of miR-7515 with FTX, and the binding of TPD52 with miR-7515, respectively. The two bindings were further validated by dual luciferase reporter assay. As a result, FTX sponged miR-7515 and miR-7515 targeted to TPD52. FTX was overexpressed in four EOC cell lines. Overexpressed FTX enhanced the cell viability, migration or invasion of EOC cells, elevated N-cadherin and TPD52 expressions, phosphorylated Met/Akt/mTOR, and inhibited E-cadherin expression. All these influences were subsequently reversed by miR-7515 mimic. Collectively, FTX regulates miR-7515/TPD52 to facilitate the migration, invasion or epithelial-mesenchymal transition of EOC through activating Met/Akt/mTOR signaling pathway.
长链非编码RNA FTX过表达与上皮性卵巢癌（EOC）患者低生存率相关，并增强肿瘤浸润。因此，我们旨在阐明尚不明确的潜在机制。应用实时定量聚合酶链反应检测FTX、miR-7515、miR-342-3p、miR-940、miR-150-5p、miR-205-5p和肿瘤蛋白D52（TPD52）的表达。利用细胞计数试剂盒-8和Transwell实验探究EOC细胞的活力、迁移或侵袭能力。进行蛋白质免疫印迹法检测E-钙黏蛋白、N-钙黏蛋白、Met、磷酸化（p）-Met、Akt、p-Akt、mTOR和p-mTOR的表达。LncBase和TargetScan分别预测miR-7515与FTX以及TPD52与miR-7515的结合。通过双荧光素酶报告基因实验进一步验证这两种结合。结果显示，FTX可吸附miR-7515，而miR-7515靶向TPD52。FTX在四种EOC细胞系中过表达。过表达的FTX增强了EOC细胞的活力、迁移或侵袭能力，提高了N-钙黏蛋白和TPD52的表达，使Met/Akt/mTOR磷酸化，并抑制E-钙黏蛋白的表达。随后，miR-7515模拟物逆转了所有这些影响。总的来说，FTX通过激活Met/Akt/mTOR信号通路调节miR-7515/TPD52，促进EOC的迁移、侵袭或上皮-间质转化。
